Project Proposal for International Science and Technology Center (ISTC)

TITLE: STUDY OF THE RED BLOOD SYSTEM, NEUROVEGETATIVE STATUS AND THE BRAIN INTEGRATIVE ACTIVITY IN ANIMALS WITH A VIEW TO REVEALING NEW CRITERIA OF BIOINDICATION OF ACUTE AND REMOTE CONSEQUENCES OF IONIZING-RADIATION EFFECTS

Summary

Basic goal of the submitted Project is the detection in laboratory animals of new diagnostic criteria and their prognostic value for the estimation of acute and remote consequences of the effect of ionizing radiation in sublethal doses, as well as development of new and perfection of the known noninvasive and inexpensive methods for mass exploration of persons of increased radiation risk. Using original complex evaluations of hematological and neurobiological studies, it is intended to determine adaptive resources of irradiated animals and their offsprings from the point of view of the degree of rehabilitation, including the biological and social value of populations. Three basic principles underlie the ideology of the Project: 1) The possibility of extrapolating the results of experimental studies for the estimation of acute and remote consequences of radiation injury and the risk rate for people, 2) Establishment of new and perfection of the known noninvasive and inexpensive criteria for the evaluation of radiation consequences, applicable for a mass exploration of people and resolution of other similar tasks, including the ecological and epidemiological ones, 3) The possibility of a maximal automatization of the used methods with a view to facilitating and standardizing mass investigations, as well as formation of data base and archives for further computer generalization of large arrays of information.
Analysis of data available in the literature and perennial experience of working in the field enable to contemplate with high probability that for the fulfillment of the task prescribed by the Project and experimental subtress of the worked out recommendations the most prospective is the application of new criteria for the estimation of acute and remote consequences of irradiation in laboratory animals. Such are, in particular, an original complex analysis of changes in the red blood system (RBS), based on the investigation of relaxation processes under the conditions of gradual reversible spherulation of erythrocytes and changes in the ability of animals to remember and learn, as well as to manifest adequate emotional motivated, aggressive and other forms of behavior, including the disturbance of the sleep-waking cycle, etc. It is intended also to investigate the neurochemical mechanisms which underlie the processes reflecting the brain integrative activity on the whole. While studying the neurobiological processes particular attention would be paid to the radiation modified natural and provoked aggressiveness which during radiation injury of animals appears to be more informative than other criteria of the brain integrative activity

The occurred and potential radionuclide disasters necessitate deepening of knowledge in the area of the biological action of ionizing radiation and estimation of the degree of risk for the health of those who had suffered and for their posterity. The Chernobyl tragedy, which, according to SRC RBE data, affected, to a considerable extent, also Western Georgia [1-5], keeps in tension millions of people in various countries. This is associated not only with the psychological tension, but also with the objective disturbances of health of a large population of people affecting their psychic and social stability. Clinical examination of a tremendous contingent and affording them a qualified assistance at the update level is an exorbitant burden placed on the economy and human resources of the countries to have suffered. If one takes into account labor-intensiveness and expensiveness of similar systematic examinations, particularly actual renders selection of the most informative noninvasive methods of check-up of population. Similar tasks are posed in the zones of ecological and social crises of various origin.

As shown by investigations, undertaken by SRC RBE, studying acute and remote consequences of radiation injury and at the experimental modelling of these processes the most informative and accessible may prove to be a complex analysis of a minimal set of hematological, cardiological and neurobiological criteria. In this connection the Project shall be realized as a unified common ideology of three sections, involving the following trends and criteria for the evaluation of acute and remote consequences of radiation injury:

Hematological criteria. Changes in the blood-forming system is known to be universal criteria of estimating the severity of radiation injury and the degree of recovery. The red blood system (RBS), however, appears not to be studied in this respect sufficiently well yet. Pursuant to recent data number of the glycophorine-A mutant erythrocytes are virtually linearly dependent on the dose received and may be used for the biological dosimentry in any period of life [6]. A model has also been made of the division of brainstem cells through the withdrawal of mutant erythrocytes [7].

Our investigations demonstrated that analysis of population spectra of PBE provides far greater diagnostically and prognostically important information than it has been known so far. In this section of the Project it is intended to create a principally new methodology on the basis of elaborated by us methods for the estimation of RBS functional state by the characteristic dynamics of changes in the PBE population spectra. The methodology relies on an original principle of investigation of relaxation processes at the erythrocyte volume variation under conditions of their gradual and reversible spherulation. The general theoretical premises, a body of mathematics and algorithmic base of the principle are described in a number of papers and in a monograph [8-15] and is also available in the SRC RBE Internet Web-site: A NEW METHOD FOR STUDYING THE RED BLOOD SYSTEM

The methodology rests on the theoretically correct and experimentally verified assumption that the families of PBE population spectra and the dynamics of alteration of the pattern of erythrocyte volumes relaxation under conditions of gradual reversible spherulation reflect adequately not only the degree of tension of functional activity of adaptive, compensatory and reparatory mechanisms of RBS, but also of the whole body. This methodology will make it possible:

1) to determine the speed and quality of the erythrocyte maturing and to evaluate the state of erythropoiesis of the pool of brainstem cell, not resorting to the exploration of bone marrow punctates and complex hematological studies.

2) to determine the rate of physiological aging of cells (biological age), the extent of functional and pathological modification (wearing) of the erythocytes' structural elements (physical) or chronological age), to estimate the rate of aging and elimination of cells having exhausted the biological resource, as well as to judge about the metabolic demands and abilities of the body in order to renovate and maintain biological full-value of the circulating and deposited erythrocyte population.

3) to estimate the functional state of the red cell membranes and oxygen density of erythrocyte hemoglobin in various age groups and judge about the rheological properties and metabolic efficiency of circulating blood.

4) to evaluate behavior of interdependence depot-circulation and define the summated reserve possibilities of adaptation mechanisms at the functional loadings on metabolism.

5) to define the RBS response at pharmacological or any other activation or depression of erythropoiesis.

6) to define radiation dose, observation period, etc dependent variation of erythrocyte mean life span.

The methodology can be widely applied not only in hematological, but also in general medical practice, including diagnostics and evaluation of efficiency of therapeutic measures in the very diverse diseases, doping control with the use of erythropoietins or hemotransfusion, for the estimation of physical condition of schoolchildren, trainage of sportsmen, professional preparation of service-men and in the case of selection of adequate representative groups - it can be applied for objective estimation of life quality of population of states and regions. The methodoly under consideration shall be realized as an integral complex of systems engineering hardware-software. Besides, on the strength of experimental findings, an experimental model was made of PBE, which will enable computer imitation of adaptive changes.

The biomedical procedure of the developed method is maximally simplified, standardized and may be combined with the routine hematological investigation, for it requires no more than 15 m l blood. The methodology may have a multipurpose application to various investigations, involving monitoring of the population life quality. In the world literature the analogous systems are not known.

Neurovegetative criteria. The vegetative nervous system is responsible for the control over all the processes of survival and subtly reacts to the variation of adaptive demands of the body depending on the changes of internal and external environment. This is reflected in the vagosympathetic balance of short-lasting (functional) and stationary character depending on the adaptation to the altered conditions of habitation environment or development of prolonged tension of regulation system during dysfunctions and pathological processes. The world scientific practice of recent years has shown that for the investigation of neurovegetative status rather effective and informative appears to be the method of analysis of heart rate variability (HRV) and respiration. General principles were elaborated for physiological interpretation of HRV data in humans and estimation of cardiological risk, including the diagnostic standards [16]. There are no analogous data for laboratory animals, although there exist separate publications devoted to these issues [17-19].

Our Center has a great experience in radiobiological investigations of this kind [20-25]. In particular, it has been demonstrated that population of rats and rabbits even of one and the same litter are not homogeneous by the vagosympathetic balance and among them as in humans one can distinctly distinguish vagotonics, normotonics and sympathicotonics. These latter appear to be far more radiosensitive, that should be taken into account while making professional selection of persons bound with radiation risk. Exposure to lethal doses of ionizing radiation elicits specific changes in the vago-sympathetic balance with a characteristic dynamics. This and a number of other data obtained by us lend support to the consideration that the investigation of variability of cardiac and respiratory rhythms turns out to be perspective for the manifestation of changes in the neurovegetative status in acute and remote periods of radiation injury.

Behavioral and neurochemical criteria. A great body of investigations deals with the effect of ionizing radiation on the CNS. The world literature, collected by us in a specialized data base, bibliographic part of which published as a separate book [26] includes more than 4 thousand papers, reviews and monographs. Nevertheless, remote neurobiological effects have been insufficiently studied yet, especially from the position of alteration of the function of brain integrity and its integrative activity. There are but a small number of works in which late changes mainly morphological, were revealed in the neural-glial architecture, the glia-vascular interrelations, as well as demyelinzation of nerve fibers, a decrease in the number of neurons, development of gliosis, etc. Dose-dependence of these changes and time of their manifestion have not been clearly defined yet. Posterity of irradiated animals have not practically been studied in this respect.

The changes detected by us in the brain integrative activity in the irradiated animals have exact neurochemical correlates in the form of characteristic alteration of quantitative content, distribution and ratio of a number of biologically active substances (mediator amino acids, biogenic amines, receptors, etc) in different brain structures and the blood serum. Morphological, including histochemical, as well as neurochemical investigations have demonstrated that in an absolute majority of animals survived after radiation these changes occur like accelerated aging and end in premature death. What kind of changes occur in the above-indicated neurobiological criteria in a remote period of radiation injury, whether they manifest themselves at the exposure to small doses, whether they affect the biological full value of irradiated animals' posterity are not known yet.

Effective use of the above-mentioned set of hematological, behavioral, neurovegetative and neurobiological criteria and a complex analysis of the results, as well as taking into account great experience of work of SRC RBE in the field, professional skill of investigators, informational maintenance and equipment with machinery, all this give grounds to believe that the Project at large will be successfully realized.

References

·  K.Nadareishvili, M.Tsitskishvili, R.Khazaradze, J.Mandjgaladze, R.Vepkhvadze, S.Kirtadze Effect of Chernobil Disaster on the Radioecological Situation in Transcaucasia. Part 2: On the Possible Manifestation of Chernobil Disaster Effect on the Health of Population in the Republic of Georgia. Radiation Studies (Tbilisi, Georgia). 1991, v. VI; pp: 152-165.

·  K.Nadareishvili, M.Tsitskishvili, G.Gachechiladze, N.Katamadze, L.Intskirveli, S.Kirtadze,J.Mandjgaladze, L.Mosulishvili, T.Sanaia, R.Khazaradze, R.Chitanava, N.Shavdia Effect of Chernobil Disaster on the Radioecological Situation in Transcaucasia. Part 1: Radionuclide Effect in Georgia. Radiation Studies (Tbilisi, Georgia). 1991, v. VI; pp: 132-151.

·  K.Nadareishvili, M.Tsitskishvili, B.Gogoladze, I.Diasamidze, G.Kvirkvinia, S.Kirtadze, L.Mosulishvili, N.Mozdokeli, A.Tordadze, R.Khazaradze, A.Chkhartishvili, N.Tsitskishvili, I.Shatberashvili, N.Shavdia, T.Shengelia, N.Katamadze The Influence of Chernobil Catastrophe on the Radioecologic State of Environment in the Caucasian Region. Part 3: The preliminary epidemiological research on radioecologic loading in the territory of Georgia after Chernobil catastrophe. Radiation Studies (Tbilisi, Georgia). 1994, v. VII; pp: 157-173.

·  K.Nadareishvili, M.Tsitskishvili, B.Gogoladze, G.Kvirkvinia, S.Kirtadze, A.Tordadze, N.Mozdokeli, R.Khazaradze, A.Chkhartishvili, N.Tsitskishvili, N.Shavdia, I.Shatberashvili, T.Shengelia The Influence of Chernobil Catastrophe on the Radioecologic State of Environment in the Caucasian Region. Part 4: Study of penetration of technogenic radionuclides in the human body via the food (alimentary) in Georgia. Radiation Studies (Tbilisi, Georgia). 1994, v. VII; pp: 174-196.

·  M.Tsitskishvili, N.Beradze, I.Diasamidze, T.Vekua, G.Gachechiladze, G.Erkomaishvili, L.Intskirveli, N.Mozdokeli, N.Katamadze, G.Kvirkvinia, M.Nozadze, A.Tordadze, A.Chkhartishvili, N.Tsitskishvili, I.Shatberashvili The Main Results on the Radioecologic Monitoring of the Caucasian Region. Part 1: Studi of Radionuclides choke up in Georgia. Radiation Studies (Tbilisi, Georgia). 1994, v. VII; pp: 197-220.

·  Kyoizumi S, Akiyama M, Cologne JB, Tanabe K, Nakamura N, Awa AA, Hirai Y, Kusunoki Y, Umeki S: Somatic cell mutations at the glycophorin A locus in erythrocytes of atomic-bomb survivors: Implications for radiation carcinogenesis. Radiation Research 146:43-52, 1996.

·  Vickers M, Beown GC, Cologne JB, Kyoizumi S. Modelling heamopoietic stem cell division by analysis of mutant red cells. Br. J. Heamatol. 2000, 110: 54-62.

·  G. Ormotsadze, K. Nadareishvili. Mechanisms of radiation injoury of red-blood cells. Proceedings of the Academy of Sciences of the Georgian SSR. Biological series. 1990, v. 138, N 1, pp 101-108..

·  G. Ormotsadze. A new approach to the research of peripherial blood erythrocytes population structure. Radiation Studies (Tbilisi, Georgia). 1994. v. 7, pp 102-112.

·  K. Nadareishvili, G. Ormotsadze, G.Grebenchuk, B.Maisuradze, B.Kulijanov. Systemotechnic complex for conductometric measuring of units dispersed in electrolyte. . Radiation Studies (Tbilisi, Georgia). 1994, v 7, pp. 312-320.

·  G. Ormotsadze, K. Nadareishvili. The Theory and Experimental Practices of Red-blood Sells Population Research. Tbilisi, 1995.

·  K. Nadareishvili, G. Ormotsadze. Study of the red-blood regulation mechanisms under physiologically normal conditions and shortly after immobilization stress. Radiation Studies (Tbilisi, Georgia). 1997, v. 8. pp.120-142.

·  G. Ormotsadze et. al. On The Possible Mechanism of Erythrocyte Ellimination From the Circulatory Bed. Georgian Medical News. 1999, N 10. pp. 35-37.

·  K.Nadareishvili, G.Ormotsadze, N.Lobzhanidze, T.Archvadze, T.Shengelia. Changes in Population Spectrum of Peripheral Blood Erythrocytes During Immobilization Stress. Bulletin of the Georgian Academy of Sciences. pp: 146-149; vol. 160; N 1; 1999.

·  G.Ormotsadze, K.Nadareishvili. New Method For Investigation of Red Blood System. Radiation Studies (Tbilisi, Georgia). 2000, v.9, pp.230-258.

·  "Heart Rate Variability - Standards of Measurement, Physiological Interpretation and Clinical Use". Special Report. Circulation, 1996, 95, 5, pp:1040-1065.

·  G.Morano, M.Grigioni, F.Tibuzzi, A.Vergari, F.Zangi. Effect of isoflurane on cardiovascular system and sympatovagal balance in New Zealand white rabbits. J.Cardiovasc. Pharmacol. 1996, 28, 4, pp:513-518.

·  V.A.Mogulecski, L.Sheil, J.Oliver, B.P.McGrayh. Power spectral analysis of heart rate variability reflects the level of cardiac autonomic activity in rabbits. J.Auton.Nerv.System. 1996, 58 (1-2), pp:18-24.

·  K.Sato, J.Satois. Circadian and short time variabilities in blood pressure and heart rate measured by telemetry in rabbits and rats. J.Auton.Nerv.Syst. 1996, 54(3), pp:235-246.

·  K.Nadareishvili, R.Gvetadze, G.Ormotsadze, I.Megreladze, A.Terknishvili, M.Khurtsia, I.Meskhishvili, M.Gvasalia, I.Maisuradze Dynamics of Cardiac Rhythm Changes During Superlethal X-Irradiation in Anesthetized by Nembutal Rats. Radiation Studies (Tbilisi, Georgia),1994, v.VII; pp:5 - 34.

·  K.Nadareishvili, I.Meskhishvili, D.Kakhiani, G.Onoprishvili Action of Signal X-Irradiation in Sublethal Doses on Heart Rate Variability (HRV) and ECG in Male Chinchilla Rabbits. Radiation Studies (Tbilisi, Georgia), 1998, v VIII; pp:27-66.

·  I.Meskhishvili, D.Kakhiani, G.Onoprishvili, G.Ormotsadze, K.Nadareishvili. Heart Rate Variability in Male Waking Chinchilla Rabbits. Bulletin of the Georgian Academy of Sciences. 1999, vol. 160; N 3; pp: 536-539;

·  I.Meskhishvil. Action of repeated X-irradiaton in sublethal doses with 3 months intervals on male chinchilla rabbits. Radiation Studies (Tbilisi, Georgia), 2000, v IX; pp 126-135.

·  I.Meskhishvil. Action of triple X-irradiaton in sublethal doses with 3 months intervals on in male chinchilla rabbits. Radiation Studies (Tbilisi, Georgia), 2000, v IX; pp:136-140

·  D.Kakhiani. Action of signal X-irradiation in lethal dose(9 Gy) on heart rate variability in male chinchilla rabbits. Radiation Studies (Tbilisi, Georgia), 2000, v IX; pp 155-141.

·  K.Nadareishvili, B.Gugushvili. Radiation Neurobiology (Bibliography). 1989, "Metsniereba", Tbilisi, Georgia.

·  M.Nokolaishvili, G.Jordanishvili, K.Nadareishvili, N.Melitauri Radiation Effect on Aggressive Rats Behavior and Quantitative Distribution of Free Amino-acids in the Rat Brain. Radiation Studies (Tbilisi, Georgia), 1991, v VI; pp: 26-37.

·  M.Nikolaishvili, K.Nadareishvili, G.Jordanishvili, N.Melitauri Superlethal X-Irradiation Effect on Aggressive Behavior and Some Neurochemical Indices in the Brain Structures of Rats. Radiation Studies (Tbilisi, Georgia), 1994, v VII; pp: 34-46.

·  M.Nikolaishvili Action of ACTH Fragment and Radiation on the Rats Aggressive Behavior and the Distribution of Transmitter Amino-acids in the Brain Structures. Radiation Studies (Tbilisi, Georgia), 2994, v VII; pp: 147-156.

·  M.Nikolaishvili, G.Jordanishvili, N.Melitauri, K.Nadareishvili The Effect of Ionizing Radiation on Aggressive Behavior in Rats, Physiological and Neurochemical Correlates. Radiation Studies (Tbilisi, Georgia), 1998, v VIII; pp: 67-78.

·  K.Nadareishvili, M.Kazakhashvili, N.Vacharadze, D.Kakhiani, I.Maisuradze, L.Tsitskishvili, G.Magradze Action of the Whole-body X-Irradiation in Sublethal and Superlethal Doses on Spontaneous and Induced Aggressive Behavior in Rats. Radiation Studies (Tbilisi, Georgia), 1998, vVIII; pp: 137-147.

·  M.Nikolaishvili. Modification of Aggressiveness, Behaviour in "Open Field" and Monoaminergic System Activity in X-Irradiated Rats. Bulletin of the Georgian Academy of Sciences. 1998, v.159, N 1; pp: 157-160.

·  M.Nikolaishvili. Radiation Effect on Aggressive Behaviour in Mice. Bulletin of the Georgian Academy of Sciences. 1998, N 2; pp: 256-259.

·  JH Schroder. Increase in aggressivnes of male mice after irradiation of paternal spermatozoa with 600 R of gama-rays as dependent on fertiliry. Behav.Genet. 1980, 10, N4, pp 387-400.